title: Biochemical and pathophysiological characterization of Helicobacter pylori asparaginase.
abstract: Asparaginase was purified from Helicobacter pylori 26695 and its pathophysiological role explored. The K(m) value of asparagine was 9.75 ± 1.81 μM at pH 7.0, and the optimum pH range was broad and around a neutral pH. H. pylori asparaginase converted extracellular asparagine to aspartate. H. pylori cells were unable to take up extracellular asparagine directly. Instead, aspartate produced by the action of the asparaginase was transported into H. pylori cells, where it was partially converted to β-alanine. Asparaginase exhibited striking cytotoxic activity against histiocytic lymphoma cell line U937 cells via asparagine deprivation. The cytotoxic activity of live H. pylori cells against U937 cells was significantly diminished by deletion of the asparaginase gene, indicating that asparaginase functions as a cytotoxic agent of the bacterium. The cytotoxic effect was negligible for gastric epithelial cell line AGS cells, suggesting that the effect differs across host cell types. An asparaginase-deficient mutant strain was significantly less capable of colonizing Mongolian gerbils. Since asparagine depletion by exogenous asparaginase has been shown to suppress lymphocyte proliferation in vivo, the present results suggest that H. pylori asparaginase may be involved in inhibition of normal lymphocyte function at the gastric niche, allowing H. pylori to evade the host immune system.
authors.affiliation: Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo, Japan. keigo@nih.go.jp
authors.forename: Keigo
authors.initials: K
authors.lastname: Shibayama
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Takeuchi
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Wachino
authors.forename: Shigetarou
authors.initials: S
authors.lastname: Mori
authors.forename: Yoshichika
authors.initials: Y
authors.lastname: Arakawa
mesh_terms: D000818:Animals; D001215:Asparaginase / Q000172:deficiency / Q000302:isolation & purification / Q000378:metabolism*; D001216:Asparagine / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D004847:Epithelial Cells; D017353:Gene Deletion; D005849:Gerbillinae; D016480:Helicobacter pylori / Q000201:enzymology* / Q000472:pathogenicity*; D006644:Histiocytes / Q000382:microbiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057131:Immune Evasion; D007700:Kinetics; D011485:Protein Binding; D014774:Virulence; D037521:Virulence Factors / Q000302:isolation & purification / Q000378:metabolism*; D015091:beta-Alanine / Q000378:metabolism
chemical_list: D037521:Virulence Factors; D015091:beta-Alanine; D001224:Aspartic Acid; D001216:Asparagine; D001215:Asparaginase